Sirtuin 6 maintains epithelial STAT6 activity to support intestinal tuft cell development and type 2 immunity

Dynamic regulation of intestinal epithelial cell (IEC) differentiation is crucial for both homeostasis and the response to helminth infection. SIRT6 belongs to the NAD+-dependent deacetylases and has established diverse roles in aging, metabolism and disease. Here, we report that IEC Sirt6 deletion leads to impaired tuft cell development and type 2 immunity in response to helminth infection, thereby resulting in compromised worm expulsion. Conversely, after helminth infection, IEC SIRT6 transgenic mice exhibit enhanced epithelial remodeling process and more efficient worm clearance. Mechanistically, Sirt6 ablation causes elevated Socs3 expression, and subsequently attenuated tyrosine 641 phosphorylation of STAT6 in IECs. Notably, intestinal epithelial overexpression of constitutively activated STAT6 (STAT6vt) in mice is sufficient to induce the expansion of tuft and goblet cell linage. Furthermore, epithelial STAT6vt overexpression remarkedly reverses the defects in intestinal epithelial remodeling caused by Sirt6 ablation. Our results reveal a novel function of SIRT6 in regulating intestinal epithelial remodeling and mucosal type 2 immunity in response to helminth infection.


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We require information from authors about some types of materials, experimental systems and methods used in many studies. Here, indicate whether each material, system or method listed is relevant to your study. If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. No statistical methods were used to predetermine sample size due to the nature of the study. Sample size determination is based on the previous experience to obtain significance and reproducibility. Basically, the sample size follows common standards employing three or more biological replicates. All sample sizes are listed in the corresponding figure legends or on the figures.

No data exclusions.
All experiments were conducted at least two times independently, and similar results were adopted for further analysis to guarantee reproducibility.
Samples were randomly allocated into the study.
This study included a lot of complicated experimental design and animal experiments, the feasibility of blinding was poor, thus blinding was not efficiently applied. The investigators were blinded to group allocation during data collection and analysis. Secondary antibody: HRP-Goat Anti-Mouse lgG (H+L) Cat: SA00001-1 Proteintech WB 1:5000 HRP-Goat Anti-Rabbit lgG (H+L) Cat: SA00001-2 Proteintech WB 1:5000 Cy3-labeled Goat Anti-Rabbit IgG (H+L) Cat: A0516 Beyotime IF 1:500 Validation of the use of Actinin antibody for mouse in WB has been provided by the manufacture's website. https://www.ptgcn.com/ products/ACTN1-Antibody-11313-2-AP.htm Validation of the use of DCLK1 antibody for mouse in IHC has been provided by the manufacture's website. https://www.abcam.com/ dcamkl1-antibody-epr6085-ab109029.html Validation of the use of FLAG antibody for mouse in WB and ChIP has been provided by the manufacture's website. https:// www.sigmaaldrich.cn/CN/zh/product/sigma/f1804 Validation of the use of Histone H3 (acetyl K56) antibody for mouse in ChIP has been provided by the manufacture's website. https:// abclonal.com.cn/catalog/A7256 Validation of the use of Ki67 antibody for mouse in IHC has been provided by the manufacture's website. https://www.abcam.com/ ki67-antibody-sp6-ab16667.html Validation of the use of MYC tag antibody in WB has been provided by the manufacture's website. https://www.ptgcn.com/products/ MYC-Antibody-60003-2-Ig.htm Validation of the use of SOCS3 antibody for mouse and human in WB has been provided by the manufacture's website. https:// www.rndsystems.com/cn/products/human-mouse-socs-3-antibody-516919_mab5696 Validation of the use of MUC2 antibody for mouse in IHC has been provided by the manufacture's website. https://www.scbt.com/ zh/p/mucin-2-antibody-h-300?requestFrom=search Validation of the use of SIRT6 antibody for mouse and human in WB has been provided by the manufacture's website. https:// www.cellsignal.cn/products/primary-antibodies/sirt6-d8d12-rabbit-mab/12486?site-search-type=Products&N=4294956287&Ntt=12486&fromPage=plp&_requestid=3909510 Validation of the use of SIRT6 antibody for mouse in IF has been provided by the manufacture's website (https://www.abcam.com/ sirt6-antibody-ab62739.html) and by Supplementary Figure 5a in our article.
Validation of the use of STAT6 antibody for mouse and human in WB has been provided by the manufacture's website. https:// www.ptgcn.com/products/STAT6-Antibody-51073-1-AP.htm Validation of the use of Phospho-STAT6 (Y641) antibody for mouse and human in WB and IF has been provided by the manufacture's website (https://www.abcam.com/stat6-phospho-y641-antibody-epr22599-78-ab263947.html) and by Supplementary Figure 5b in our article.
Validation of the use of -Tubulin antibody for mouse and human in WB has been provided by the manufacture's website. https:// www.ptgcn.com/products/TUBA1B-Antibody-11224-1-AP.htm Validation of the use of CD3 FITC antibody for mouse in flow cytometry has been provided by the manufacture's website. https:// www.biolegend.com/en-us/products/fitc-anti-mouse-cd3-antibody-45 Validation of the use of CD4 PE/Cyanine7 antibody for mouse in flow cytometry has been provided by the manufacture's website. https://www.biolegend.com/en-us/products/pe-cyanine7-anti-mouse-cd4-antibody-1919 Validation of the use of CD11b PE antibody for mouse in flow cytometry has been provided by the manufacture's website. https:// www.biolegend.com/en-us/products/pe-anti-mouse-human-cd11b-antibody-349 Validation of the use of CD11c PE antibody for mouse in flow cytometry has been provided by the manufacture's website. https:// www.biolegend.com/en-us/products/pe-anti-mouse-cd11c-antibody-1816 Validation of the use of CD16/32 antibody for mouse in flow cytometry has been provided by the manufacture's website. https:// www.biolegend.com/en-us/products/trustain-fcx-anti-mouse-cd16-32-antibody-5683 Validation of the use of CD19 PE antibody for mouse in flow cytometry has been provided by the manufacture's website. https:// www.biolegend.com/en-us/products/pe-anti-mouse-cd19-antibody-1530 nature portfolio | reporting summary Note that full information on the approval of the study protocol must also be provided in the manuscript.
The study did not involve field-collected samples.
Xinxiang Medical University, IACUC